在荧光漂白恢复测量中荧光恢复起始时间的研究

在荧光漂白恢复测量中,漂白过程结束的准确时间相对于漂白控制信号后沿会有一定的延迟时间,不易确定.光电倍增管快门开启又要推迟一段时间.因此,荧光恢复起始时间,即漂白结束的瞬间不能准确给出,开始阶段数据收集不全.本文分析了荧光恢复起始时间不准确所造成的误差,测量了起始时间.并尽量减少此误差,使恢复初期收集到的数据更完全,荧光恢复过程的时间座标更准确.

THE STUDY ON INITIAL TIME OF FLUORESCENCE RECOVERY IN THE FPR EXPERIMENT

In fluorescence photobleaching recovery (FPR) , delay time exists between the first moment while the signal is sent out to close the bleaching shutter and the second moment while the shutter acts really. In order to protect the PMT from being damaged by the intensive light, the PMT shutter should wait for a moment to open (100-200ms) in general. The moment, when the bleaching is completed or the fuorescence recovery begins, couldn't be exactly determined. The fluorescence intensity at t=0 (i.e.f(0)) could not be detected accurately as well. These make the acquired data inaccurate in t-axis, and reduced the accuracy of result. For rapid diffusion, it becomes more serious.In this paper, we have estimated the errors arisen from the initial recovery time, and analysed the factors which affect the delay time of bleaching shutter. Futhermore, the delay time has been measured and the time error has been corrected. After appropriate selection of PMT waiting time as short as possible, the data are acquired more completely during the initial stage of fluorescence recovery, i,e.F(0) is more precise.